Stain for crustacean cuticle?


Hi histonetters,
a colleague and I are planning to stain the cuticle oflive
copepods (small marine crustaceans). We are still looking for the appropriate 
stain and I wonder if you have any good ideas. There are different components 
in the cuticle layers of the copepods that we could stain. These are:
- chitin (C8H13O5N)n: Polyacetylglukosamin
- layer of polymerized protein (arthrodin) and lipoids
- resilin: rubber-like protein, couldn't find more specific description
- surface coat: mucopolysaccharides and carbohydrates
- protein-chitin complexes (a protein called arthropodin)
To summarize, the components in the cuticle are proteins, water, chitin and 
lipoids. The ideal stain would need to
fulfill the following requirements:
- be non-toxic as it will be applied on life specimens
- be photostable as the animals will remain in aquaria for a while, and
preserved samples will have to be stored for at least a few weeks after
the experiment
- stain the exosceleton (see above) and NOT too much of the tissue underneath
- be bacterial resistant, i.e. not degradable by marine bacteria
- be non-ambiguous, meaning the wavelengths it should not interfere
with some naturally occurring fluorescence (which we actually don't know
yet) or that of formaldehyde or other preservatives
- be stable in formaldehyde or alcohol as the specimens will be
preserved after the experiment
- be affordable.
Could you recommend something?
Also, I wonder if any of you know the autofluorescence spectra of formaldehyde 
and methanol (70%) or know where I could look this up.
Bodil B

Bodil Bluhm
Institute of Marine Science
School of Fisheries and Ocean Sciences
University of Alaska Fairbanks
P.O. Box 757220
245 O'Neill Bld (for shipping)
Fairbanks AK 99775-7220
Phone: 907 474 7407
Fax: 907 474 7204

<< Previous Message | Next Message >>